Changes of respiration rate and fatty acid composition in cell membranes of pupae of the large cabbage white, Pieris brassicae (Lepidoptera: Pieridae) during winter

Document Type : Paper, Persian

Author

Abstract

Cell membranes are very sensitive to low temperatures and undergo changes and adaptations to prevent this damage. One of the most common changes is the increase in unsaturated to saturated fatty acids ratio of phospholipids, which leads to increase the fluidity of membranes at low temperatures. In this study, overwintering pupae of the large cabbage white, Pieris brassicae were collected during November 2014 to April 2015. To investigate the metabolic quiescent, changes in respiratory gases (O2 and CO2) and respiration rate (‍CO2/O2) were measured using GC. After extracting the whole body lipids, phospholipids were separated from other lipids by solid phase extraction (SPE) and then fatty acids and their changes were investigated using the GC. Respiration rate decreased about 30% from 0.9 in November to 0.6 in February and March, indicating that metabolism has reached to its lowest level during these period and insect has been in deep pupal diapause. Two 16-carbon and four 18-carbon fatty acids were identified in cell membranes. During winter, the highest decrease was observed in two saturated fatty acids C16: 0 and C18: 0 and the highest increase occurred in C16: 1. In general, unsaturated /saturated fatty acids ratio increased about 40 percent during the winter months. The results indicated that in spite of decreasing metabolism level in the whole body, in order to enhance the cold-hardiness, dynamic changes occurred in cell membranes, allowing insect to adapt successfully to winter hard conditions.

Keywords


Atapour, M. (2016) Investigation of Overwintering and Cold Tolerance in Pupae of Large Cabbage White, Pieris brassicae (Lepidoptera: Pieridae) in Iran. Iranian Journal of Plant Protection Science 46(2), 269-276. [In Persian with English summary].
Atapour, M. (2017) Cryoprotectants in Lab-reared and Overwintering Pupae of Large
Cabbage White, Pieris brassicae (Lepidoptera: Pieridae).  Iranian Journal of Plant
Protection Science
48 (1), 139-150. [In Persian with English summary].
Atapour, M., Moharramipour, S. & Barzegar, M. (2007) Seasonal changes of fatty acid compositions in overwintering larvae of rice stem borer, Chilo suppressalis
(Lepidoptera: Pyralidae). Journal of Asia-Pacific Entomology 10, 33-38.
Bahrndorff, S., Petersen, S.O., Loeschcke, V. Overgaard, J. & Holmstrup, M. (2007) Differences in cold and drought tolerance of high arctic and sub-arctic populations of Megaphorura arctica Tullberg 1876 (Onychiuridae: Collembola). Cryobiology 55, 315-323.
Bashan, M. & Cakmak, O. (2005) Changes in composition of phospholipid and
triacylglycerol fatty acids prepared from prediapausing and diapausing individuals of Dolycoris baccarum and Piezodorus lituratus (Heteroptera: Pentatomidae). Annals of Entomological Society of America 98, 575-579.
Bennett, V. A. & Lee, R. E. (1997) Modeling seasonal changes in intracellular
freeze-tolerance of fat body cells of the gall fly Eurosta solidaginis. Journal of
Experimental Biology
200, 185-192.
Choi, G. T., Casu, M. & Gibbons, W. A. (1993) N.m.r. lipid profiles of cells, tissues and body fluids. Neutral, non-acidic and acidic phospholipid analysis of Bond Elut
chromatographic fractions. Biochemical Journal, 290 (3), 717-721.
Danks, H. V. (1987) Insect Dormancy: an Ecological Perspective. Biological Survey of Canada, Ottawa, 439 pp.
Danks, H. V., Kukal, O. & Ring, R. A. (1994) Insect cold hardiness-insights from the arctic. Arctic 47, 391-404.
Ding, L. Li, Y. & Goto, M. (2003) Physiological and biochemical changes in summer and winter diapause and non-diapause pupae of the cabbage armyworm, Mamestra
brassacae
L. during long-term cold acclimation. Journal of Insect Physiology 49, 1153-1159.
Folch, J., Lees, M. & Sloane-Stanley, G. H. (1967) A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry 226, 497-509.
Goto, M., Sekine, Y., Outa, H., Hujikura, M. & Suzuki, K. (2001) Relationships between cold hardiness and diapause, and between glycerol and free amino acid contents in
overwintering larvae of the oriental corn borer, Ostrinia furnacalis. Journal of Insect Physiology 47, 157-165.
Irwin, J. T. & Lee, R. E., Jr. (2003) Cold winter microenvironments conserve energy and improve overwintering survival and potential fecundity of the goldenrod gall fly,
Eurosta solidaginis. Oikos 100, 71-78.
Jõgar, K., Metspalu, L., Hiiesaar, K., Luik, A., Martin, A.-J., Mänd, M., Jaaniso, R. & Kuusik, A. (2005) Physiology of diapause in pupae of Pieris brassicae L. (Lepidoptera: Pieridae). Agronomy Research 3(1), 21-37.
Kaneko, J. & Katagiri, C. (2006) A simple method to discriminate diapause from
non-diapause pupae in large and small white butterflies, Pieris brassicae and P. rapae crucivora. Naturwissenschaften 93(8), 393-396.
Kayukawa, T., Chen, B., Hoshizaki, S., & Ishikawa, Y. (2007) Upregulation of a
desaturase is associated with the enhancement of cold hardiness in the onion maggot, Delia antiqua. Insect Biochemistry and Molecular Biology 37, 1160-1167.
Khani, A., Moharramipour, S., Barzegar, M. & Naderi-Manesh, H. (2007) Comparison of fatty acid composition in total lipid of diapause and non-diapause larvae of Cydia pomonella (Lepidoptera: Tortricidae). Insect Science, 14: 125-131.
Kostal, V. & Simek, P. (1998) Changes in fatty acid composition of phospholipids and
triacylglycerides after cold acclimation of an aestivating insect prepupa. Journal of Comparative Physiology (B) 168, 453–460.
Kostal, V., Shimada, K. & Hayakawa, Y. (2000) Induction and development of winter larval diapause in a drosophilid fly, Chymomyza costata. Journal of Insect Physiology 46, 417-428.
Metcalf, L. C., Schmitz, A. A. & Pelka, J. R. (1966) Rapid preparation of methyl esters from lipid for gas chromatography analysis. Analytical Chemistr38, 514-515.
Michaud, M. R. & Denlinger, D. (2006) Oleic acid is elevated in cell membranes during rapid cold-hardening and pupal diapause in the flesh fly, Sarcophaga crassipalpis.
Journal of Insect Physiology 52(10), 1073-1082.
Michaud, M. R. & Denlinger, D. L. (2007) Shifts in the carbohydrate, polyol, and amino acid pools during rapid cold-hardening and diapause-associated cold-hardening in flesh flies (Sarcophaga crassipalpis): a metabolomic comparison. Journal of Comparative Physiology B: Biochemical, Systemic, and Environmental Physiology 177, 753-763.
Moreau, R., Olivier, D., Gourdoux, L. & Dutrieu, J. (1981) Carbohydrate metabolism in Pieris brassicae L. (Lepidoptera); variations during normal and diapausing
development. Comparative Biochemistry and Physiology 68 (B), 95-99.
Overgaard, J., Tomčala, A., Sørensen, J.G., Holmstrup, M. Henning Krogh, P. Šimek, P. & KOŠTÁL, V. (2008) Effects of acclimation temperature on thermal tolerance and membrane phospholipid composition in the fruit fly Drosophila melanogaster. Journal of Insect Physiology 54(3), 619-629.
Pullin, A. S. & Bale, J. S. (1989) Influence of diapause and temperature on cryoprotectant synthesis and cold hardiness in pupae of Pieris brassicae. Comparative Biochemistry and Physiology (A) 94, 499-503.
Rozsypal, J., Koštál, V., Berková, P., Zahradníčková, H. & Šimek, P. (2014) Seasonal changes in the composition of storage and membrane lipids in overwintering larvae of the codling moth, Cydia pomonella. Journal of Thermal Biology 45, 124–133.
Saito, A., K. Koga, & B. Sakaguchi. (1982) Changes in polysome content during
development after diapause of Bombyx mori embryos. Federation of European
Biochemical Societies 150(2), 449-453.
Shimuzu, I. (1992) Comparsion of fatty acid composition in lipids of diapause and
non-diapause eggs of Bombyx mori. Comparative Biochemistry and Physiology (B) 102,713-716.
Sinensky, M. (1974) Homeoviscous adaptation; a homeostatic process that regulates the
viscosity of membrane lipids in Escherchia coli. Proceedings of the National Academy of Sciences, USA 71(2), 522- 525.
Singer, S. J. & Nicolson, G. L. (1972) The fluid mosaic model of the structure of cell
membranes. Science 175, 720-731.
Starling, A. P., East, J. M. & Lee, A. G. (1993) Effects of phosphatidylcholine fatty acyl chain length on calcum binding and other functions of the [Ca 2+ - Mg 2+]-ATPase. Biochemistry 32, 1593-1600.
Stola, A., Tollarova, M., Overgaard, J., Simek, P. & Kostal, V. (2006) Seasonal 
acquisition of chill tolerance and restructuring of membrane glycerophospholipids in an overwintering insect: triggering by low temperature, desiccation and diapause 
progression. Journal of Experimental Biology 209, 4102-4111.
Vukašinović, E.L., Pond, D.W., Worland, M.R., Kojić, D., Purać, J., Popović, Ž.D. & rey, K. B. & Storey, J. M. (1988) Freeze tolerance in animals. Physiological Reviews 68, 27-84.
Thompson, A.C. & F.M. Davis. (1981) The effect of temperature on the rate of metabolism of lipids and glycogen in diapausing southwestern corn borer, Diatraea grandiosella. Comparative Biochemistry and Physiology  (A) 70(4), 555-558.
TomcaGrubor-Lajšić, G. (2015) Diapause induces remodeling of the fatty acid composition of 
membrane and storage lipids in overwintering larvae of Ostrinia nubilalis, Hubn.
(Lepidoptera: Crambidae). Comparative Biochemistry and Physiology (B) 184, 36-43. doi: 10.1016/j.cbpb.2015.02.003.
Vukašinović, E.L., Pond, W.D., Worland, M.R., Kojić, D., Purać, J., Blagojević, P.D. & Grubor-Lajšić, G. (2013) Diapause induces changes in the composition and
biophysical properties of lipids in larvae of the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae). Comparative Biochemistry and Physiology (B) 165, 219–225.